Fatty acids, coco, esters with 3,3'-oxybis[1,2-propanediol]

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-03-14 to 2017-08-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

yes

Remarks:

threshold concentration as limit concenrtation (see overall remarks)

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Qualifier:

according to guideline

Guideline:

other: OECD No 23; Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures

Qualifier:

according to guideline

Guideline:

other: OECD 126 “Short Guidance on the threshold approach for acute fish toxicity"

Version / remarks:

2010

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Duplicate samples from the freshly prepared test media filtrate of the only test concentration and the control were taken at the start of the test.
For the determination of the stability of the test item under the test conditions and of the maintenance of the test item concentrations during the test period, duplicate samples from the test media of the only test concentration and the control were collected at the end of the test (after 96 hours) from the approximate centre of the aquaria.
Additionally, one sample of the freshly prepared stock solution filtrate was taken prior to test start for analysing the test item concentration in the filtrate.

Details on test solutions:

Test Concentration: Geometric mean measured Concentration: 18.53 mg test item/L and a control.
The lowest EC50 value of existing relevant algae (growth rate) or acute invertebrate (daphnia) tests is set as threshold concentration. This threshold approach was performed in order to demonstrate that the test item has no toxic effects on the test fish up to the threshold concentration or at 100 mg (active ingredient)/L.
Control: In the control, test water was used without addition of the test item.
Dosage of Test Item: A stock solution with a concentration of 70 mg test item/L was prepared by mixing 699.0 mg test item in 9985.7 ml test water by stirring for 48 hours. Then, the undissolved test item was separated by filtration (0.65 and 0.45 µm cellulose acetate filter). The test medium was analysed. According to the pre-analysis the filtrate was not further diluted. The test medium was prepared just before introduction of the test fish (= start of the test).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species: Zebrafish (Danio rerio)
Size and Weight: Juveniles; The mean body length of the fish* in the test was 2.15 cm ± 0.18 cm (Mean ± SD), the mean body wet weight 0.080 g ± 0.016 g (Mean ± SD).
* 10 fish from the test fish batch were measured before the start of the test

Sex: Male and female
Origin: The zebrafish were obtained from our in house the brood stock of ibacon GmbH.
Holding Conditions: All fish were obtained and held in the laboratory for at least 12 days before the start of the test. They were held in water of the quality to be used in the test for at least seven days immediately before testing under the following conditions:
Light: 16 hours photoperiod daily
Temperature: 21- 25 °C
Oxygen concentration: at least 80 % of the air saturation value
Feeding: three times per week or daily until 24 hours before the test was started During the last 7 days prior to the start of the test no fish (0 %) died in the test fish batch. Therefore the mortalities in the fish batch were below 5 % and the fish batch was accepted.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

no

Hardness:

2.5 mmol/L (= 250.0 mg/L) as CaCO3

Test temperature:

22.9 to 23.4 °C

pH:

7.4 to 8.0

Dissolved oxygen:

94 to 100 % % of the air saturation value

Nominal and measured concentrations:

Geometric mean measured Concentration: 18.53 mg test item/L and a control.
The lowest EC50 value of existing relevant algae (growth rate) or acute invertebrate (daphnia) tests was set as threshold concentration. This threshold approach was performed in order to demonstrate that the test item has no toxic effects on the test fish up to the threshold concentration or at 100 mg (active ingredient)/L.

Details on test conditions:

Test Environment: Controlled environmental room
Measurement of pH-Values, Dissolved Oxygen and Water Temperature:
The water temperature, pH-values and the dissolved oxygen concentrations were determined daily in the test media of each treatment group.
Water Temperature: 22.9 to 23.4 °C
pH-Values of the Test Water: 7.4 to 8.0
Dissolved Oxygen Concentrations:
94 to 100 % % of the air saturation value
Light Regime: 16 h light : 8 h dark;
30 min dawn/dusk period was provided
Light Intensity: 1050 to 1070 lux
Aeration of the Test Water: The test media were slightly aerated during the test.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 18.53 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

In the control and the only test geometric mean measured test concentration of 18.53 mg test item/L, all fish survived until the end of the experiment and showed no sublethal effects during the exposure time.
96-hour LC50: > 18.53 mg test item/L (geometric mean measured)
95 % Confidence Interval: n.d.
96-hour LC20: > 18.53 mg test item/L (geometric mean measured)
95 % Confidence Interval: n.d.
96-hour LC10: > 18.53 mg test item/L (geometric mean measured)
95 % Confidence Interval: n.d.
96-hour LC0: ≥ 18.53 mg test item/L (geometric mean measured)
96-hour LC100: > 18.53 mg test item/L (geometric mean measured)
96-hour NOEC: ≥ 18.53 mg test item/L (geometric mean measured)
96-hour LOEC: > 18.53 mg test item/L (geometric mean measured)

Reported statistics and error estimates:

No statistical analysis was performed.
The LC50 could not be quantified due to the absence of toxicity of the test item and was determined to be higher than the tested concentration. The NOEC and the LOEC were determined directly from the raw data.

Validity criteria fulfilled:

yes

Remarks:

Control: no fish died during the test; Dissolved Oxygen Concentration: in the test media did not fall below 94 % of air saturation value during the test

Conclusions:

Derived effect concetration: 96h-LC50 (Danio rerio) > 18.53 mg/L (geometric mean measured)

Executive summary:

The purpose of this study was to evaluate the acute toxicity of the test item Hostastat FE 20 LIQ to fish. For this purpose, juvenile Zebrafish were exposed in a static test to the only concentration under defined conditions for 96 hours. The recorded effects were the mortality and sublethal effects on the fish. A threshold approach was pursued. The test method of application and the test system are recommended by the test guidelines and Zebrafish is one of the recommended test species. This threshold approach was performed in order to demonstrate that the test item has no toxic effects on the test fish up to the threshold concentration or at 100 mg (active ingredient)/L. The lowest EC50 value of existing relevant algae or acute invertebrate (e.g. daphnia) tests was set as threshold concentration. In the interest of animal welfare and efficient use of resources, it is important to avoid the unnecessary use of animals whenever possible. In the field of aquatic toxicology, this especially applies to the acute toxicity testing of fish according to OECD 203. The threshold approach described hereafter addresses fish toxicity by using a single-concentration test (limit test) requiring less fish compared to the full acute fish toxicity study. It is based on the derivation of a threshold concentration from relevant algae and acute invertebrate (e.g. daphnia) tests at which fish toxicity is tested. As no mortality occurred in the limit test using the threshold concentration, the threshold concentration can be used as a surrogate LC50 value in the further hazard or risk assessment.

Guidelines:

Commission Regulation (EC) No 440/2008, Annex, Part C, C.1.: "Acute Toxicity for Fish", Official Journal of the European Union, May 30, 2008

OECD Guideline for Testing of Chemicals, Section 2, No. 203, "Fish, Acute Toxicity Test", adopted July 17, 1992

OECD Series on Testing and Assessment, No. 23, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures", December 15, 2000

OECD Guidance Document, Series on Testing and Assessment No. 126, “Short Guidance in the Threshold Approach for Acute Fish Toxicity”, May 31, 2010

SANCO/3029/99 rev.4 11/07/00: Residues: Guidance for generating and reporting methods of analysis in support of pre-registration data requirements for Annex II (part A; Section 4) and Annex III (part A; Section 5) of directive 91/414

Material and Methods

Test Item: Hostastat FE 20 LIQ; batch no.: ESD0021912; purity: 100 % (w/w), according to certificate of analysis

Test Species: Juvenile Zebrafisch (Danio rerio) mean length: 2.15 cm ± 0.18 cm; the zebrafish were obtained from our in house the brood stock of ibacon GmbH.

Test Design: This study encompassed two treatment groups (one geometric mean measured test item concentration of 18.53 mg test item/L and one control) each containing 7 individuals. The acute toxicity to unfed juvenile Zebrafish was determined in an aerated, static, 96-hour test. The test fish were observed at test start and after approximately 2, 24, 48, 72 and 96 hours test duration for sublethal effects and mortality.

Endpoints: NOEC after 96 h, LOEC after 96 h; LC50: lethal concentration producing 50 % mortality after 96 h of exposure.

Test Concentration: Geometric mean measured Concentration: 18.53 mg test item/L and a control.

The lowest EC50 value of existing relevant algae (growth rate) or acute invertebrate (daphnia) tests was set as threshold concentration. This threshold approach was performed in order to demonstrate that the test item has no toxic effects on the test fish up to the threshold concentration or at 100 mg (active ingredient)/L.

Test Conditions: Water temperature: 22.9 to 23.4 °C; pH value: 7.4 to 8.0; dissolved oxygen concentration: 94 to 100 % % of the air saturation value; photoperiod: 16 h light - 8 h dark; light intensity: 1050 to 1070 lux and thus were within the ranges requested by guideline OECD 203.

Results

Biological test results: In the control and the only test geometric mean measured test concentration of 18.53 mg test item/L, all fish survived until the end of the experiment and showed no sublethal effects during the exposure time.

Analytical test results

The quantification of the test item Hostastat FE 20 LIQ in the test samples was performed by analysing the Total Organic Carbon content (TOC).At the start of the test 119 % of the nominal test concentration in the filtrate was found (average of the only test concentration). After 96 hours test duration, 94 % of the nominal value was determined (average of the only test concentrations). During the test the fish were exposed to a mean of 106 % of nominal. All reported results refer to geometric mean measured concentrations.

Conclusion: The toxic effect of the test item Hostastat FE 20 LIQ to Zebrafish (Danio rerio) was assessed in a static limit test. Based on the test results the 96-hour LC50 was determined to be > 18.53 mg test item/L based on geometric mean measured concentrations. The NOEC was determined to be ≥ 18.53 mg test item/L also based geometric mean measured concentrations.

This study was classiefied acceptable and satisfied the guideline requirements for fish acute test studies.

Description of key information

96h-LC50 (Danio rerio) > 18.53 mg/L; 96h-NOEC > 18.53 mg/L (geometric mean measured) (threshold (limit) test, OECD 203, 2017)

Note: The key value was inserted in order to enable the application of the PNEC calculator. The inserted concentration is higher than the effect concentration used as basis for the PNEC derivation and therefore does not bias the PNEC calculation performed by the PNEC calculator.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

18.53 mg/L

Additional information

In a reliable key study, acute toxicity of the submission substance to fish was evaluated according to OECD 203 and in compliance with GLP (RL1). For this purpose, juvenile Zebrafish were exposed to a threshold test concentration for 96 hours in a static test (limit test). A threshold approach was performed in order to demonstrate that the submission substance has no toxic effects on fish up to a threshold concentration, i.e. the lowest derived EC50 value from existing relevant algae or acute invertebrate (e.g. daphnia) tests. The threshold concentration was verified by analysing the Total Organic Carbon content (TOC). A control was run in parallel.

No mortality or sublethal effects were observed in neither treatment group nor control. The 96-hour LC50 was determined to be > 18.53 mg test item/L based on geometric mean measured concentrations. The NOEC was determined to be ≥ 18.53 mg test item/L also based geometric mean measured concentrations.

This study was classified acceptable and satisfied the guideline requirements for fish acute test studies. In conclusion, the test was considered reliable and adequate for the environmental hazard assessment for aquatic organisms.